Precise and dynamic measurement of intracellular metabolite levels has been hampered by difficulties in differentiating between adsorbed and imported fractions and the subcellular distribution between cytosol, endomembrane compartments and mitochondria. In the present study, genetically encoded FRET (Förster resonance energy transfer)-based sensors were deployed for dynamic measurements of free cytosolic glucose and ATP with varying external supply and in glucose-transport mutants. Moreover, by using the FRET sensors in a microfluidic platform, we were able to monitor in vivo changes of intracellular free glucose in individual yeast cells. We demonstrate the suitability of the FRET sensors for gaining physiological insight by demonstrating that free intracellular glucose and ATP levels are reduced in a hxt5Δ hexose-transporter mutant compared with wild-type and other hxtΔ strains.
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Research Article|
November 12 2010
Dynamic analysis of cytosolic glucose and ATP levels in yeast using optical sensors
Clara Bermejo;
Clara Bermejo
1
1Department of Plant Biology, Carnegie Institution for Science, 260 Panama Street., Stanford, CA 94305, U.S.A.
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Farzad Haerizadeh;
1Department of Plant Biology, Carnegie Institution for Science, 260 Panama Street., Stanford, CA 94305, U.S.A.
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Hitomi Takanaga;
Hitomi Takanaga
1Department of Plant Biology, Carnegie Institution for Science, 260 Panama Street., Stanford, CA 94305, U.S.A.
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Diane Chermak;
Diane Chermak
1Department of Plant Biology, Carnegie Institution for Science, 260 Panama Street., Stanford, CA 94305, U.S.A.
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Wolf B. Frommer
Wolf B. Frommer
3
1Department of Plant Biology, Carnegie Institution for Science, 260 Panama Street., Stanford, CA 94305, U.S.A.
3To whom correspondence should be addressed (email wfrommer@stanford.edu).
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Publisher: Portland Press Ltd
Received:
July 06 2010
Revision Received:
September 09 2010
Accepted:
September 20 2010
Accepted Manuscript online:
September 20 2010
Online ISSN: 1470-8728
Print ISSN: 0264-6021
© The Authors Journal compilation © 2010 Biochemical Society
2010
Biochem J (2010) 432 (2): 399–406.
Article history
Received:
July 06 2010
Revision Received:
September 09 2010
Accepted:
September 20 2010
Accepted Manuscript online:
September 20 2010
Citation
Clara Bermejo, Farzad Haerizadeh, Hitomi Takanaga, Diane Chermak, Wolf B. Frommer; Dynamic analysis of cytosolic glucose and ATP levels in yeast using optical sensors. Biochem J 1 December 2010; 432 (2): 399–406. doi: https://doi.org/10.1042/BJ20100946
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